PMC:7573174 / 14638-30512 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/7573174","sourcedb":"PMC","sourceid":"7573174","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7573174","text":"Enzymatic supramolecular biomaterials\nThe noncovalent and enzymatic nature of EISA leads to the use of enzymes, which is ubiquitous in biological systems, to trigger self-assembly to generate different molecular ensembles. This approach, directly connecting the transformation of biomaterials with numerous physiological and pathological processes, has received considerable exploration in the past two decades. By discussing the following representative examples, we briefly introduce the applications of enzymatic supramolecular biomaterials in (i) tissue engineering, (ii) cancer therapy, (iii) drug delivery, (iv) imaging, and (v) other applications.\n\nTissue engineering\nExtracellular matrix (ECM), a three-dimensional network of extracellular biomolecules, plays important roles in cell adhesion, proliferation, differentiation, and communication (Clark and Brugge, 1995[13]; Engler et al., 2006[18]; Lauffenburger and Horwitz, 1996[47]; Sternlicht and Werb, 2001[74]). The highly dynamic feature of ECM coincides with EISA, which provides an insight into developing ECM mimicking biomaterials. A recent study reports the design of supramolecular phosphoglycopeptides (sPGPs) assemblies that transform 2D cell sheets into 3D cell spheroids, mimicking the functions of ECM (Wang et al., 2017[88]). Based on the ligand-receptor interactions between phosphopeptides (5) and glycopeptides (6), these two components self-assemble to form nanoparticles, which transform into nanofibers upon dephosphorylation in the cell milieu (Figure 2A(Fig. 2); References in Figure 2: Feng et al., 2018[23]; Wang et al., 2019[84]). The co-culture of cancer cells (Saos-2) and stromal cells (HS-5) in different ratios is able to tailor the level of ALP expression, which controls the morphological transformation of the two-components assemblies for enabling cell morphogenesis or for inducing cell death (Wang et al., 2019[84]). In this case, the supramolecular assemblies act as a context-dependent signal for controlling cell fates in the co-culture that mimics tumor microenvironment. This insight leads to the development of a homotypic precursor that achieves spatiotemporal control of noncovalent interactions for mimicking ECM dynamics (Wang et al., 2019[85]). In a different study reported by Qi et al. (2018[69]), incorporating proangiogenic drug into ECM mimicking hydrogel facilitates vasculogenesis because the densely glycosylated hydrogel serves as a reservoir for the sustainable release of drug and induces capillary morphogenesis both in vitro and in vivo (Qi et al., 2018[69]).\nBesides extracellular biomimetic nanomaterials, supramolecular assemblies are able to mimic intracellular biomacromolecules for enzyme sequestration. For example, a phosphopeptide precursor (7) targeting ER undergoes dephosphorylation and forms nanofibers of 8 that sequestrate enzymes enriched in ER (Figure 2B(Fig. 2)) (Feng et al., 2018[23]). A trimethylated phosphopeptide undergoes ALP-instructed intracellular morphological transition to form bundles of artificial filaments inside the cells (Feng et al., 2020[21]). The biomimetic filaments hardly interfere with endogenous compartments, providing insights for intracellular filaments engineering.\n\nCancer therapy\nCancer is the second leading cause of death in the 21st century. Although various treatments have been developed, a large number of patients suffer from cancer progression and recurrence. Recent studies have revealed the complexity of cancer, such as tumor heterogeneity, tumor microenvironment, multidrug resistance (MDR), abnormal metabolism, and epigenetic alteration (Hanahan and Weinberg, 2011[31]). These findings not only explain the failure of chemotherapies in certain cancers to some extent, but also demand new strategies to improve treatment efficacy. For example, connecting taurine to D-peptides via ester bonds boosts their cellular uptake, which is able to counteract the increased efflux pump and the decreased drug absorption caused by MDR (Zhou et al., 2015[123], 2018[120]). Integrating drugs with peptides that are responsive to enzymes upregulated on or inside cancer cells is a useful approach to stabilize some prodrugs until they reach the target sites. Furin-controlled Arg-Val-Arg-Arg-Cys(StBu)-Lys(taxol)-2-cyanobenzothiazole (CBT-Taxol) condensation (Yuan et al., 2015[109]), ALP-instructed self-assembly of anti-cancer peptide NapGDFDFpYSV (Gao et al., 2019[26]), and CES-guided release of coumarin (Zhang et al., 2019[112]) are a few representative examples of this approach.\nBesides making up important parts in prodrugs, the supramolecular assemblies of small molecules are emerging as anticancer drug candidates (Kuang et al., 2014[44]; Kuang and Xu, 2013[46]; Yang et al., 2007[108]). The nanofibers of peptide derivatives formed pericellularly or intracellularly can induce cell death, cytoskeletal rearrangement, and compromised membrane integrity (Li et al., 2018[50]). Specifically, a self-assembling motif (e.g. Nap-FF) is conjugated with an enzyme-responsive (ALP, CES, ENTK or β-galactosidase) substrate to generate a precursor. Upon encountering cancer cells that overexpress these enzymes, the precursor transforms to the hydrogelator via enzymatic bond cleavage, followed by self-assembling process to form nanofibers. By tuning the self-assembling abilities, these nanofibrils can be either innocuous or cytotoxic, exhibiting a molecular dynamic continuum for cancer therapy (Kuang et al, 2014[44]; Feng et al., 2017[20]). For example, a C-terminal methylated phosphopeptide (9) is a nontoxic precursor at its optimal concentration. However, 9 induces cell stress and shows strong synergism with NF-κB, verified by adding BAY 11-7085 (BAY) to make the cells sensitive to stress (Figure 3A(Fig. 3); References in Figure 3: He et al., 2020[32]; Wang et al., 2016[82]) (Zhou et al., 2018[121]). A CES cleavable peptide precursor exhibits similar synergism with cisplatin for inhibiting drug-resistant ovarian cancer cells (Li et al., 2015[52]).\nBeing the substrates of the overexpressed ALP in some cancer cell lines (e.g., HeLa and Saos-2), several phosphopeptide precursors are able to form toxic nanofibrils on or inside the cancer cells, causing membrane insertion (Du et al., 2019[15]), efficient and selective cell removal (Kuang et al., 2017[45]), cancer cell inhibition (Feng et al., 2019[19], 2020[24]; Wang et al., 2016[83], 2019[87]), or cell rupture (Li et al., 2017[49]). As reported by Zhang et al., a ruthenium-based phosphopeptide trimer (10) that favors fast and complete dephosphorylation by glycosylphosphatidylinositol-anchored placental alkaline phosphatase (GPI-anchored PLAP) in lipid raft (Figure 3B(Fig. 3)) enables the cell rupture. Specifically, the assemblies form patches on HeLa cells, resulting in reinforced focal adhesion (FA) on filopodia. The opposite direction of cell migration generates mechanical stress that eventually causes cell rupture.\nAside from targeting an enzyme or a protein, cell type-specific, dual-targeting nanomedicines offer a better chance for successful targeting cancer cells without inducing acquired drug resistance. Common combinations of dual targets include enzymes and receptors (Wang et al., 2019[92]), enzymes and ligands (Wang et al., 2016[82]), two different enzymes (Zheng et al., 2019[117]), and simply two drugs with different targets (Mang et al., 2019[63]). The dual-targeting concept is highly useful in designing compounds to control cell fates (Zhou et al., 2017[125]), including targeting downregulation in cancer cells (Feng et al., 2017[25]).\nOwing to the precise spatiotemporal control of EISA, targeting subcellular organelles or subcellular proteins with EISA is an emerging field in cancer therapy (Feng et al., 2018[22]; He et al., 2018[36], 2020[33]). Integrating extracellular EISA with the mitochondrial targeting ability of triphenyl phosphonium, the peptide derivative (11) induce mitochondrial dysfunction and the release of cytochrome c, therefore killing the cancer cells (Figure 3C(Fig. 3)) (Wang et al., 2016[82]). Incorporating ENTK to induce perimitochondrial self-assembly directly delivers chloramphenicol (CLRP) into mitochondria, selectively sensitizing and killing cancer cells (He et al., 2020[33]). A phosphopeptide bearing AVPI segment, which is the antagonistic motif to the inhibitors of apoptotic proteins (IAPs), sequestrates IAPs and selectively induce apoptosis of cancer cells (He et al., 2020[34]). Containing Flag-tag and acting as the substrate of ENTK, a negatively charged peptide derivative (12) traffics a nuclear protein, histone protein (H2B) to mitochondria (Figure 3D(Fig. 3)). Being the first example of EISA to traffic endogenous proteins, this work provides insights to understand inter-organelle cross talk (He et al., 2020[32]).\n\nDrug delivery\nIn recent years, many efforts have focused on incorporating enzymatic supramolecular hydrogels in drug delivery systems because it is easy to synthesize the hydrogels that are biocompatible and versatile. Aiming for desired therapeutic effects, common goals of drug delivery are (i) improving pharmacokinetic properties, (ii) involving active targeting, and (iii) achieving sustained release.\nDue to the hydrophobic and unspecific nature of many anticancer drugs, clinical administration of these chemotherapies often suffers from low solubility, poor bioavailability, and severe side effects. Commercially available formulations address these problems by encapsulating drugs into liposomes, such as Doxil, or proteins, such as Abraxane. Alternatively, the enzyme-responsive feature of EISA provides another route by selective targeting of enzymes overexpressed on/in cancer cells. In particular, the morphology of precursor changes upon enzymatic cleavage, which results in self-assembly to form drug-encapsulating hydrogels with enhanced cellular internalization and thus improved drug efficacy. For example, a self-delivery hydrogel of LND-GFFpY, reported by Wu et al. (2019[94]), exhibits a 560-fold increase in solubility compared with lonidamine (LND) alone, as well as selective killing of cancer cells. Similarly, a drug-peptide amphiphile comprising a p53-activating peptide ligand, PMI, reported by Yang et al., enhances cellular uptake and achieves nuclear accumulation (Liang et al., 2020[57]). Besides, the CES modified silicon nanoparticles, which respond specifically to CES substrates, can form nanogels around the particles. The unique core-shell structure serves as a nanocarrier for doxorubicin (DOX) (Wu et al., 2018[93]).\nIn addition to better pharmacokinetics, supramolecular hydrogels, due to their slow biodegradation, function as local depots for sustained release of drug. For instance, two MMP-9 responsive peptide amphiphiles, which form spherical aggregates in water, undergo morphological transition to form fibers encapsulating DOX. As reported by Kalafatovic et al. (2016[41]), the fibrous networks serve as depots at the invasive border characterized by MMP-9 overexpression Choosing the substrates responsive to different enzymes is able to finely tune the gelation location. In a study reported by Cheng et al. (2019[12]), a cathepsin B (CTSB) responsive prodrug forms intracellular nanofibers for preventing local tumor recurrence after surgery. Intracellular ALP-responsive co-assembly of dexamethasone (Dex) phosphate and a hydrogelator precursor Nap-Phe-Phe-Tyr(H2PO3)-OH boosts the anti-inflammation efficacy of Dex, as shown by Tang et al. (2017[75]). They also reported similar efficacy enhancement when Dex is covalently conjugated to a homotypic precursor (Tang et al., 2018[76]), or replaced by other drugs like etoposide (Kiran et al., 2018[43]).\nAnother strategy is to use enzyme-responsive gold nanoparticles to enhance photothermal therapy (PTT) efficacy. Hu et al. (2017[39]) conjugated gold nanoparticles with two types of peptide segments on the surface, one is an MMP-2 substrate peptide (CPLGVRGDDRGD), and the other is a self-assembling peptide (CKKKLVFF). Being cut by MMP-2, the strengthened H-bonding between intermolecular self-assembling peptides leads to the aggregation of nanoparticles and red shift in localized surface plasmon resonance (LSPR), thus enhanced PTT efficiency (Figure 4(Fig. 4)) (Hu et al., 2017[39]). A subsequent study by Wang et al. shows that ALP-responsive gold nanoparticles exhibit similar effects (Yang et al., 2018[103]).\n\nImaging\nBeing in situ and noninvasive, medical imaging techniques such as magnetic resonance imaging (MRI), near-infrared (NIR) fluorescence, computed tomography (CT), positron tomography (PET) and photoacoustic (PA) imaging lead to the development of molecular imaging in the past two decades. The tunability and biocompatibility of EISA allow aberrant levels of enzymes to catalyze the accumulation of imaging agents in situ to obtain accurate and precise imaging. Pioneered by Liang et al. (2010[58]), furin-guided CBT condensation initiates self-assembly process to form nanoparticles with dense imaging agents, facilitating MicroPET imaging (Wang et al., 2019[78]; Liu et al., 2015[62]), fluorescence (Li et al., 2019[55]), and phosphorescence imaging (Li et al., 2018[51]) in different cases. As reported by Liang et al., ALP-instructed self-assembly generates nanofibers for enhanced CT (Zheng et al., 2016[119]) and MRI (Dong et al., 2017[14]). Spatiotemporal tuning allows the assemblies to form either extracellularly for cell delineation (Zhao et al., 2017[116]), or intracellularly for imaging (Wu et al., 2018[95]; Wang et al., 2019[90]). Acting on precursors that are responsive to other significant enzymes in physiologic processes, EISA can monitor membrane dynamics. For example, the conjugation of cholesterol with phosphopeptide generates 13 to target lipid raft (Figure 5A(Fig. 5); References in Figure 5: Wang et al., 2018[79]; Zheng et al., 2016[118]). After dephosphorylation by ALP within the lipid raft, the probe selectively assembles in plasma membrane, precisely showing membrane dynamics for an extended period (Figure 5B(Fig. 5)) (Wang et al., 2018[79]). EISA also finds applications in monitoring other dynamic processes or significant components, such as autophagy (Lin et al., 2017[60][61]), Caspas-1 (Li et al., 2016[54]), CTSB (Ni et al., 2019[67]), and fibroblast activation protein-α (FAP-α) (Zhao et al., 2019[115]).\nThe dual targeting strategy, that is, the use of dual targeting motif (either ligands or enzymes), also applies in molecular imaging (Chen et al., 2019[9]; Hai et al., 2019[30]; An et al., 2020[3]). As reported by Zheng et al. (2016[118]), a precursor (14) is able to differentiate the extra- and intracellular environment based on the dual targeting motifs that react with both extracellular ALP and intracellular glutathione (GSH), generating different orders of assemblies (15 and 16) (Figure 5C(Fig. 5)). It is also a common strategy to use multiple imaging agents in the precursor. Such a combination can provide comprehensive information to the question of interest, as evidenced by the synergism of NIR and MRI for in vivo imaging, as elegantly demonstrated by Yan et al. (2019[100]). Another approach is to incorporate molecular imaging with therapeutic agents to conduct diagnosis and therapy at the same time. The conjugation of drugs with imaging agents provides rich opportunities to develop theranostics (Yuan et al., 2019[110]) that improve imaging process and drug efficacy (Zhang et al., 2018[111]; Ji et al., 2018[40]).\n\nOther applications\nApart from a large number of applications for cancer diagnosis and treatment, biomaterials based on noncovalent interactions also play important roles in neuronal development (Mei et al., 2015[65]), antibody production (Wang et al., 2019[91]), and would healing (Li et al., 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